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DNA translocation to giant unilamellar vesicles during electroporation is independent of DNA size
Delft Univ Technol, Dept Chem Engn, van der Maasweg 9, NL-2629 HZ Delft, Netherlands..
Delft Univ Technol, Dept Chem Engn, van der Maasweg 9, NL-2629 HZ Delft, Netherlands..ORCID iD: 0000-0003-2240-4260
Delft Univ Technol, Dept Chem Engn, van der Maasweg 9, NL-2629 HZ Delft, Netherlands..
Delft Univ Technol, Dept Chem Engn, van der Maasweg 9, NL-2629 HZ Delft, Netherlands..
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2019 (English)In: Soft Matter, ISSN 1744-683X, E-ISSN 1744-6848, Vol. 15, no 45, p. 9187-9194Article in journal (Refereed) Published
Abstract [en]

Delivery of naked DNA molecules into living cells via physical disruption of the membrane under electric pulses has potential biomedical applications ranging from gene electro-transfer, electro-chemotherapy, to gene therapy, yet the mechanisms involved in DNA transport remain vague. To investigate the mechanism of DNA translocation across the cell membrane, giant unilamellar vesicles (GUVs) were electroporated in the presence of DNA molecules keeping the size of the DNA molecules as a variable parameter. We experimentally determined the translocation efficiency for each size of the DNA molecule, to compare the results with the existing and conflicting theories of the translocation mechanism i.e. stochastic threading and bulk electrophoresis. We observed that the translocation efficiency is independent of DNA size (ranging from 25-20 000 bp, bp = base pairs), implying that DNA molecules translocate freely across the electro-pores in the lipid membrane in their native polymer conformation, as opposed to unravelling and threading through the electro-pore. Bulk electrophoretic mobility determines the relationship between translocation efficiency and the size of the DNA molecule. This research provides experimental evidence of the mechanistic understanding of DNA translocation across lipid membranes which is essential for devising efficient and predictable protocols for electric field mediated naked DNA delivery.

Place, publisher, year, edition, pages
ROYAL SOC CHEMISTRY , 2019. Vol. 15, no 45, p. 9187-9194
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Biophysics
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URN: urn:nbn:se:kth:diva-266422DOI: 10.1039/c9sm01274eISI: 000502302700021PubMedID: 31595286Scopus ID: 2-s2.0-85075636816OAI: oai:DiVA.org:kth-266422DiVA, id: diva2:1387778
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Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20200122

Available from: 2020-01-22 Created: 2020-01-22 Last updated: 2020-01-22Bibliographically approved

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